Crystalline 1-methylcarbapenem derivatives

ABSTRACT

This invention provides crystalline forms of a 1-methylcarbapenem derivative of formula (I) or of pharmaceutically acceptable salts thereof.  
                 
 
     The crystalline forms of the 1-methylcarbapenem derivative exhibit excellent antibiotic activity against various bacterial strains and sufficient stability for practical use.

BACKGROUND OF THE INVENTION

[0001] 1. Field of the Invention

[0002] This invention is directed to crystalline forms of1-methylcarbapenem derivatives or of pharmaceutically acceptable saltsthereof which exhibit excellent antibiotic activity against variousbacterial strains and are stable enough to keep for a long time. Thisinvention is directed compositions for the prevention or treatment ofbacterial infections containing a crystalline form of the presentinvention as an active ingredient. This invention is directed to uses ofa crystalline form of the present invention in order to prepare amedicament for the prevention or treatment of bacterial infections. Thisinvention is directed to methods for the preventing or treatingbacterial infections which comprise administering to a warm-bloodedanimal in need of such prevention or treatment an effective amount of acrystalline form of the present invention. Further this invention isdirected to processes for the preparation of crystalline forms of thepresent invention.

[0003] 2. Description of Related Art

[0004] The 1-methylcarbapenem derivative of formula (I) is disclosed inJapanese Patent Application Publication Hei-10-204086 and Hei-11-071277and in U.S. Pat. No. 6,090,802 (corresponds to Hei-10-204086). Thiscompound (I) exhibits excellent antibiotic activity not only againstGram-positive bacterial strains but also against Gram-negative bacterialstrains and can be expected to become a useful antibiotic agent.However, the compound (I) prepared according to the Example of JapanesePatent Application Publication Hei-11-071277 (which is Example 72 ofU.S. Pat. No. 6,090,802) was obtained by lyophilization as anon-crystalline powder. This powder is unstable and is a materialdifficult to keep for a long time. There are many problems in practicaluse of the powder as a medicament, especially as an antibiotic agent.The inventors made many efforts in order to solve these problems andhave found that certain crystalline forms of compound (I) are extremelystable compared to the non-crystalline powder of compound (I) and areuseful medicaments, especially, practically useful antibiotic agents.The stable crystalline forms of this invention include a crystallineform of compound (I) ½ carbonate containing ½ ethanol (I-1), acrystalline form of compound (I) containing ½ ethanol (I-2), acrystalline form of compound (I) and a crystalline form of compound (I)containing ¼ ethanol and {fraction (3/2)} water (I-3).

BRIEF DESCRIPTION OF THE INVENTION

[0005] This invention is directed to

[0006] 1. a 1-methylcarbapenem derivative of formula (I) or apharmaceutically acceptable salt thereof in crystalline form,

[0007] 2. a 1-methylcarbapenem derivative of formula (I-1) incrystalline form,

[0008] 3. a 1-methylcarbapenem derivative of formula (I-2) incrystalline form,

[0009] 4. a 1-methylcarbapenem derivative of formula (I) in crystallineform,

[0010] 5. a 1-methylcarbapenem derivative of formula (I-3) incrystalline form,

[0011] 6. pharmaceutical compositions for the prevention or treatment ofbacterial infections containing a crystalline form of a1-methylcarbapenem derivative according to any one of 1 to 5 as anactive ingredient,

[0012] 7. uses of a crystalline form of a 1-methylcarbapenem derivativeaccording to any one of 1 to 5 in order to prepare a medicament for theprevention or treatment of bacterial infections, and

[0013] 8. methods for preventing or treating bacterial infections whichcomprise administering to a warm-blooded animal in need of suchprevention or treatment an effective amount of a crystalline form of a1-methylcarbapenem derivative according to any one of 1 to 5.

DETAILED DESCRIPTION

[0014] Carbapenem derivatives of formula (I) are disclosed in U.S. Pat.No. 6,090,802 and Japanese Patent Application Publications Hei-10-204086and Hei-11-071277, and exhibit potent activity against Gram-positive andGram-negative bacterial strains.

[0015] Carbapenem derivatives of formula (I) can exist aspharmaceutically acceptable salts. The term “a pharmaceuticallyacceptable salt” as used herein and in the claims is intended to includesalts which are usually able to be used as medicaments.

[0016] The compound of formula (I) has basic groups such as a tertiaryamino group and a guanidino group and can be converted to apharmaceutically acceptable acid addition salt when treated with anappropriate acid employing conventional techniques. Such acid additionsalts include inorganic acid salts such as hydrochlorides,hydrobromides, sulfates and phosphates; organic acid salts such ascarbonates, acetates, benzoates, oxalates, maleates, fumarates,tartrates and citrates; and sulfonates such as methanesulfonates,benzenesulfonates and p-toluenesulfonates.

[0017] The compound of formula (I) has an acidic group such as acarboxyl group and can be converted to a pharmaceutically acceptablebase addition salt when treated with an appropriate base employingconventional techniques. Such base addition salts include alkali metalsalts such as sodium salts, potassium salts and lithium salts; alkalineearth metal salts such as calcium salts and magnesium salts; metal saltssuch as aluminum salts, iron salts, zinc salts, copper salts, nickelsalts, and cobalt salts; and quaternary ammonium salts such as ammoniumsalts.

[0018] When allowed to stand in the air, certain forms of the compound(I) and pharmaceutically acceptable salts thereof absorb or adsorb waterand can form hydrates. In certain cases forms of the compound (I) andpharmaceutically acceptable salts thereof absorb certain solvents andcan form solvates. The compound (I) of this invention andpharmaceutically acceptable salts thereof include such hydrates andsolvates. Such salts, hydrates and solvates are preferably sodium salts,hydrochlorides, sulfates, carbonates, hydrates or solvates of ethanol;most preferably carbonates, hydrates or solvates of ethanol.

[0019] The compound of formula (I-1) represents the ½ ethanol solvate ofthe ½ carbonate salt of the 1-methylcarbapenem derivative of formula(I). The compound of formula (I-2) represent the ½ ethanol solvate ofthe 1-methylcarbapenem derivative of formula (I). The compound offormula (I-3) represents the {fraction (3/2)} hydrate and ¼ ethanolsolvate of the 1-methylcarbapenem derivative of formula (I).

[0020] Crystalline Form:

[0021] The crystalline forms of the present invention are solids whichhave regular arrangements of atoms (group of atoms) in three-dimensionalstructure and repeat the arrangements. The crystals are different froman amorphous solid that has no such regular arrangement of atoms in athree-dimensional structure.

[0022] In general, certain compounds produce a plurality of crystallineforms (polymorphic crystals) according to crystallization conditions,crystals of which are different in their three-dimensional arrangementof atoms and in physicochemical properties. This invention may includeeach of such crystalline forms and mixtures no less than two thereof.The crystalline form of the 1-methylcarbapenem derivative of formula(I-1) shows main peaks at interplanar spacings d=6.65, 5.68, 4.86, 4.57and 4.03 Å in the X-ray powder diffraction pattern obtained with a CuK_(α) irradiation of λ=1.54 Å. The main peaks have intensities not lessthan 74, which is the relative intensity when the intensity of the peakat 4.57 Å is evaluated as 100.

[0023] The crystalline form of the 1-methylcarbapenem derivative offormula (I-2) shows main peaks at interplanar spacings d=10.57, 7.12,5.34, 5.23, 4.91 and 4.26 Å in the X-ray powder diffraction patternobtained with a Cu K_(α) irradiation of λ=1.54 Å. The main peaks haveintensities not less than 56, which is the relative intensity when theintensity of the peak at 4.91 Å is evaluated as 100.

[0024] The crystalline form of the 1-methylcarbapenem derivative offormula (I) shows main peaks at interplanar spacings d=8.07, 5.08, 4.89,4.44, 4.39 and 4.19 Å in the X-ray powder diffraction pattern obtainedwith a Cu K_(α) irradiation of λ=1.54 Å. The main peaks have intensitiesnot less than 48, which is the relative intensity when the intensity ofthe peak at 5.08 Å is evaluated as 100.

[0025] The crystalline form of the 1-methylcarbapenem derivative offormula (I-3) shows main peaks at interplanar spacings d=7.02, 4.90,4.64, 4.59 and 4.03 Å in the X-ray powder diffraction pattern obtainedwith a Cu K_(α) irradiation of λ=1.54 Å. The main peaks have intensitiesnot less than 65, which is the relative intensity when the intensity ofthe peak at 7.02 Å is evaluated as 100.

[0026] Preparation:

[0027] The compound of formula (I) can be prepared by the same techniqueas described, or by a similar procedure to that described in U.S. Pat.No. 6,090,802, Japanese Patent Application Publication Hei-10-204086 andHei-11-071277.

[0028] The crystalline forms of this invention were obtained, forexample,

[0029] 1) by dissolution of compound (I) or a pharmaceuticallyacceptable salt thereof in an appropriate solvent which can readilydissolve it,

[0030] 2) by, if necessary, concentration of the solution, addition tothe solution of an appropriate solvent which can slightly dissolvecompound (I) or a pharmaceutically acceptable salt thereof or coolingthe solution in order to obtain a supersaturated solution and hence tocrystallization, and

[0031] 3) by isolation of the crystals and then drying of the crystals.Precipitation of the crystals begins spontaneously in the vessel, orprecipitation can also begin or be accelerated by addition ofcrystalline seeds or by mechanical stimulations such as ultrasonic waveirradiation and scratching on the surface of the vessel.

[0032] Pharmaceutically acceptable salts of compound (I) are preferablyhydrochlorides, sulfates and carbonates; most preferably carbonates. Thepharmaceutically acceptable salts can be prepared by addition ofnecessary amount of a desired acid or base to a solution of compound(I).

[0033] When solutions of the compound (I) or pharmaceutically acceptablesalts thereof are treated, the solutions of these compounds are usuallytreated between 0 and 60° C. in order to avoid decomposition of thesecompounds.

[0034] The preferred temperature of crystallization of these compoundsis between 0 and 10° C.

[0035] Methods of concentration of solutions of the compound (I) orpharmaceutically acceptable salts thereof are an evaporation methodusing a rotary evaporator under reduced or normal pressure upon heatingand a concentration method using a reverse osmotic membrane. The reverseosmotic membrane used in concentration of an aqueous solution can beselected from polyacrylonitrile membranes, polyvinyl alcohol membranes,polyamide membranes and cellulose acetate membranes.

[0036] Examples of solvents which can readily dissolve compound (I) orpharmaceutically acceptable salts thereof are water, dimethyl sulfoxide,dimethylformamide and methanol, preferably water.

[0037] Examples of solvents which can slightly dissolve compound (I) orpharmaceutically acceptable salts thereof are C₂-C₄ alcohols such asethanol, propanol and butanol; ketones such as acetone and methyl ethylketone; ethers such as diethyl ether and tetrahydrofuran; and esterssuch as methyl acetate and ethyl acetate; preferably ethanol andacetone; most preferably ethanol.

[0038] The starting compound (I) which is isolated as a lyophilizedpowder can be used. A crude reaction solution containing compound (I)can also be used because it is possible to purify by crystallization.

[0039] Supersaturation can be accomplished by concentration of anaqueous solution of compound (I) at between 30 and 60° to a saturatedaqueous solution, followed by gradually cooling to between 0 and 10° C.or accomplished by gradual addition of an appropriate solvent which canslightly dissolve compound (I) or pharmaceutically acceptable saltsthereof, such as ethanol or acetone, to the saturated aqueous solution,if necessary, followed by cooling.

[0040] Crystalline forms of this invention preferably precipitate whenaqueous solutions of compound (I) or pharmaceutically acceptable saltsare concentrated, if necessary, followed by the addition of a solventwhich can slightly dissolve these compounds, followed by cooling. Morepreferably crystals of this invention precipitate when aqueous solutionsof compound (I) or pharmaceutically acceptable salts thereof areconcentrated, if necessary, followed by the addition of ethanol oracetone and then cooling.

[0041] Most preferably, the preferred crystalline form of compound (I-1)precipitates when an aqueous solution of compound (I) is concentrated,followed by saturation with carbon dioxide, addition of ethanol andcooling; the preferred crystalline form of compound (I-2) precipitateswhen an aqueous solution of compound (I) is concentrated, followed bythe addition of ethanol and by cooling (preferably by irradiation withultrasonic waves); the preferred crystalline form of compound (I)precipitates when an aqueous solution of compound (I) is concentrated,followed by cooling; the preferred crystalline form of compound (I-3)precipitates when an aqueous solution of compound (I) is concentrated,followed by addition of ethanol and by cooling.

[0042] The precipitated crystals are isolated, for example, byfiltration, centrifugation or decantation. If necessary, the isolatedcrystals can be washed with an appropriate solvent. Preferably thecrystals are washed at first with the solvent which is used incrystallization, and then washed with a solvent such as ethanol,acetone, and ether.

[0043] The isolated crystals are dried at between 10 and 50° C.,preferably at between 20 and 30° C. until the weight of the crystalsbecome constant. If necessary, they may be dried in the presence ofdrying agents such as silica gel and calcium chloride under reducedpressure.

[0044] Utility:

[0045] The crystalline forms of compound (I-1), (I-2), (I) and (I-3) areeasy to use practically as a medicament and are extremely stablecompared to the lyophilized powder of compound (I) which is disclosed inJapanese Patent Application Publication Hei-11-071277.

[0046] The crystalline forms of this invention exhibit a wide spectrumof antibiotic activity and potent antibacterial activities againstGram-positive and Gram-negative strains and anerobic bacteria, as wellas bacteria producing cephalosporinase. When the antibacterialactivities of the crystals of this invention were determined by theagar-plate dilution method, they exhibited potent antibacterialactivities against various bacteria, for example, Gram-positive strainssuch as Staphylococcus aureus, methicillin-resistant Staphylococcusaureus, Streptococcus pneumoniae, Enterococcus and the like;Gram-negative strains such as Escherichia coli, Bacillus dysenteriae,Klebsiella pneumoniae, Proteus vulgaris, Serratia, Enterobacteriaceae,Pseudomonas aeruginosa and the like; and anerobic bacteria such asbacteroides fragilis. The crystalline forms of this invention exhibitedpotent antibacterial activity against Helicobacter pylori which is oftendetected in the patients with chronic gastritis and peptic ulcers.

[0047] When appropriate solutions of the crystalline forms of thisinvention were administered to mice, they exhibited long half-valueperiods of blood concentration and good urinary recovery compared tothose of similar compounds known to those skilled in the art.

[0048] When the crystalline forms of this invention were subcutaneouslyadministered to mice infected systemically with Staphylococcus aureus,Streptococcus pneumoniae, Escherichia coli or Psuedomonas aeruginosa,they exhibited excellent treatment effect. The crystalline forms of thisinvention, therefore, are useful medicaments (especially antibacterialagents).

[0049] When the crystalline forms of this invention are used as amedicament (especially as an antibacterial agent) they can beadministered alone or as a mixture of said crystalline forms of thisinvention and a pharmaceutically acceptable excipient(s) and diluent(s);they can be administered in various dosage forms such as tablets,capsules, granules, powders, or syrups for oral administration, such asinjections for parenteral administration or such as ointments fortopical application.

[0050] Such dosage forms are prepared by methods known to those skilledin the art using additives such as excipients, binders, disintegrants,lubricants, stabilizers, corrigents, suspending agents, diluents,solvents for formulation, assisting agents for dissolution, and topicalanaesthetic agents.

[0051] Examples of excipients include sugar derivatives such as lactose,sucrose, glucose, mannitol and sorbitol; starch derivatives such as cornstarch, potato starch, α starch, dextrin and carboxymethylstarch;cellulose derivatives such as crystalline cellulose, low-substitutedhydroxypropylcellulose, hydroxypropylmethylcellulose,carboxymethylcellulose and internally-cross-linked sodiumcarboxymethylcellulose; arabic gum; dextran; pululan; silicatederivatives such as light silicic acid anhydride, synthetic aluminumsilicate and magnesium aluminate metasilicate; phosphate derivativessuch as calcium phosphate; carbonate derivatives such as calciumcarbonate, and sulfate derivatives such as calcium sulfate.

[0052] Examples of binders include excipients as described above;gelatin; polyvinylpyrrolidone; and macrogol.

[0053] Examples of disintegrants include excipients as described above,and chemically modified starch and cellulose derivatives such as sodiumcross-carmelose, sodium carboxymethylstarch and cross-linkedpolyvinylpyrrolidone.

[0054] Examples of lubricants include talc; stearic acid; metal stearatederivatives such as calcium stearate and magnesium stearate; colloidalsilica; bee gum; waxes such as bee's wax and spermaceti; boric acid;glycol; carboxylic acid derivatives such as fumaric acid and adipicacid; sodium carboxylate derivatives such as sodium benzoate; sulfatederivatives such as sodium sulfate; leucine; lauryl sulfate derivativessuch as sodium lauryl sulfate and magnesium lauryl sulfate; silicic acidderivatives such as silicic acid anhydride and silicic acid hydrate; andstarch derivatives as described for the excipients.

[0055] Examples of stabilizers include para-oxybenzoic acid esters suchas methylparaben and propylparaben; alcohols such as chlorobutanol,benzyl alcohol, and phenethyl alcohol; benzalkonium chloride; phenolderivatives such as phenol and cresol; thimerosal; acetic anhydride, andsorbic acid.

[0056] Examples of corrigents include sweetening, souring and flavoringagents all of which are usually used.

[0057] Examples of solvents for formulation include water, ethanol, andglycerin.

[0058] Examples of assisting agents for dissolution include non-ionicsurfactants, and anionic surfactants.

[0059] Examples of topical anaesthetic agents include lidocainehydrochloride, and mepivacaine hydrochloride.

[0060] Dosage forms for oral administration include, for example, soliddosage forms such as tablets, coated tablets, capsules, troches,powders, fine granules, granules, and dry syrups and liquid dosage formssuch as syrups. Dosage forms for parenteral administration include, forexample, injections, dripping infusions and suppositories. Dosage formsfor topical application include, for example, ointments, tinctures,creams, and gels.

[0061] Preferable dosage forms of the crystalline 1-methylcarbapenemderivatives of this invention are injections and dripping infusions.Suitable dosage levels for the crystalline forms depend on the age, bodyweight, and symptoms of the patient and are usually from 10 mg(preferably 50 mg) to 6000 mg (preferably 4000 mg) for an adult humanper day, which dosage can be administered as a single dose or dividedinto several doses throughout the day.

EXAMPLES

[0062] The following examples, reference examples, test examples andformulation examples further illustrate this invention.

[0063] All NMR spectra in examples and reference examples weredetermined in deuterated water using tetramethylsilane or in anothersolvent using sodium 3-(trimethylsilyl)propionate as internal standards,respectively.

[0064] Throughout the chemical structures the following abbreviationsare used with the following meaning:

[0065] PNB : 4-nitrobenzyl

[0066] PNZ: 4-nitrobenzyloxycarbonyl

Example 1

[0067](1R,5S,6S)-2-[(2S,4S)-2-[(3S)-3-(2-Guanidinoacetylamino)pyrrolidin-1-ylcarbonyl]-1-methylpyrrolidin-4-ylthio]-6-[(1R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylicAcid·½ Carbonate·½ Ethanol

[0068] To a solution of 4-nitrobenzyl(1R,5S,6S)-2-[(2S,4S)-2-[(3S)-3-[2-[2,3-bis(4-nitrobenzyloxycarbonyl)guanidino]acetylamino]pyrrolidin-1-ylcarbonyl]-1-methylpyrrolidin-4-ylthio]-6-[(1R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate(9.4 g) in a mixture of tetrahydrofuran (235 ml) and water (140 ml) wasadded 7.5% palladium on carbon (9.4 g, which contains water (53.1%)) andthe resulting mixture was stirred under a hydrogen atmosphere at 35° C.for 2 hours. At the end of this time the catalyst was removed byfiltration and the filtrate was washed with ether. The ether andtetrahydrofuran were evaporated in vacuo and the resulting residue waschromatographed on a reverse phase column (Cosmosil 75C 18PREP (trademark) manufactured by Nacalai tesque Inc.) using a mixture ofacetonitrile and water as the eluant. The fractions containing thedesired product were combined and concentrated to approximately 50 ml invacuo. Ethanol (100 ml) and dry ice were added to the concentrate andthe resulting solution was allowed to stand in an ice bath. Theresulting precipitate was filtered and washed successively with amixture of ethanol and water (2:1), ethanol and ether to afford thetitle compound as colorless crystals (3.15 g). Melting point: 228-233°C. (dec)

[0069] Infrared spectrum (KBr)ν max cm⁻¹:3331, 2968, 2875, 2791, 1755,1669, 1637, 1453, 1386, 1339, 1312, 1283, 1254.

[0070] NMR spectrum (400 MHz, D₂O) δ ppm: 1.13-1.24 (4.5H, m), 1.30 (3H,d, J=6.4 Hz), 1.57-1.72 (1H, m), 1.93-2.10 (1H, m), 2.15-2.35 (1H, m),2.27, 2.29 (3H, s×2), 2.68-2.88 (2H, m), 3.09 (1H, d, J=10.6 Hz),3.29-3.73 (7H, m), 3.75-3.93 (2H, m), 4.01 (2H, s), 4.12-4.30 (2H, m),4.38-4.50 (1H, m).

[0071] Elemental analysis: calculated for C₂₃H₃₅N₇O₆S·½H₂CO₃·½C₂H₆OCal.: C 49.73%; H 6.64%; N 16.57%; S 5.42%; Found: C 49.57%; H 6.86%; N16.68%; S 5.47%

[0072] A X-ray powder diffraction pattern of the crystalline productshown in FIG. 1 was obtained with a Cu K_(α) irradiation of λ=1.54 Å.The vertical axis of the x-ray powder diffraction pattern indicates thediffraction intensity in units of counts/second (CPS). The horizontalaxis indicates the diffraction angle as the value 2θ. The interplanarspacing d can be calculated using the equation 2d sin θ=nλ in which n is1.

Example 2

[0073] (1R,5S,6S)-2-[(2S,4S)-2-[(3S)-3-(2-Guanidinoacetylamino)pyrrolidin-1-ylcarbonyl]-1-methylpyrrolidin-4-ylthio]-6-[(1R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylicAcid·½ Ethanol

[0074] To a solution of 4-nitrobenzyl(1R,5S,6S)-2-[(2S,4S)-2-[(3S)-3-[2-[2,3-bis(4-nitrobenzyloxycarbonyl)guanidino]-acetylamino]pyrrolidin-1-ylcarbonyl]-1-methyl-pyrrolidin-4-ylthio]-6-[(1R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate(10.00 g) in a mixture of tetrahydrofuran (250 ml) and water (150 ml)was added 7.5% palladium on carbon (10.00 g, which contains water(53.1%)) and the resulting mixture was stirred under a hydrogenatmosphere at 35° C. for 2 hours. At the end of this time the catalystwas removed by filtration, the filtrate was washed with ether and wasfiltered through a membrane filter. The resulting filtrate wasconcentrated to approximately 50 ml in vacuo. To the concentrate wasadded ethanol (100 ml) and the resulting mixture was irradiated withultrasonic waves in order to precipitate crystals, and then allowed tostand in an ice bath. The precipitated crystals were filtered and washedsuccessively with a mixture of ethanol and water (2:1), ethanol andether and then dried to afford the title compound as colorless crystals(3.30 g).

[0075] Melting point: 235-250° C. (dec)

[0076] Infrared spectrum (KBr)ν max cm⁻¹: 3405, 3344,3273, 3207, 2969,2883, 2795, 1760, 1673, 1644, 1591, 1553, 1452, 1415, 1381, 1370, 1341,1311, 1283, 1255.

[0077] NMR spectrum (400 MHz, D₂O) δ ppm: 1.15-1.25 (4.5H, m), 1.30 (3H,d, J=6.4 Hz), 1.57-1.72 (1H, m), 1.93-2.13 (1H, m), 2.15-2.35 (1H, m),2.27, 2.29 (3H, s×2), 2.68-2.88 (2H, m), 3.08 (1H, d, J=10.7 Hz),3.29-3.73 (7H, m), 3.75-3.93 (2H, m), 4.01 (2H, s), 4.16-4.31 (2H, m),4.37-4.49 (1H, m).

[0078] Elemental analysis : calculated for C₂₃H₃₅N₇O₆S·½C₂H₆O Cal.: C51.41%; H 6.83%; N 17.49%; S 5.72%; Found: C 51.13%; H 6.96%; N 17.17%;S 5.72%

[0079] A X-ray powder diffraction pattern of the crystalline productshown in FIG. 2 was obtained with a Cu K_(α) irradiation of λ=1.54 Å.The vertical axis of the x-ray powder diffraction pattern indicates thediffraction intensity in units of counts/second (CPS). The horizontalaxis indicates the diffraction angle as the value 2θ. The interplanarspacing d can be calculated using the equation 2d sin θ=nλ in which n is1.

Example 3

[0080](1R,5S,6S)-2-[(2S,4S)-2-[(3S)-3-(2-Guanidinoacetylamino)pyrrolidin-1-ylcarbonyl]-1-methylpyrrolidin-4-ylthio]-6-[(1R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylicAcid·½ Ethanol

[0081] To a solution of 4-nitrobenzyl(1R,5S,6S)-6-[(1R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-1-methyl-2-[(3S)-3-[2-[3-(4-nitrobenzyloxycarbonyl)guanidino]-acetylamino]pyrrolidin-1-ylcarbonyl]pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate(112 mg) in a mixture of tetrahydrofuran (2.2 ml) and water (2.2 ml) wasadded 7.5% palladium on carbon (112 mg, which contains water (53.1%))and the resulting mixture was stirred under a hydrogen atmosphere at 35°C. for 2 hours. At the end of this time the catalyst was removed byfiltration and the filtrate was washed with ether and was filteredthrough a membrane filter. The filtrate was concentrated toapproximately 1 ml in vacuo. To the resulting concentrate was addedethanol (2 ml) and the mixture was irradiated with ultrasonic waves inorder to precipitate crystals, and then allowed to stand in an ice bath.The precipitated crystals were filtered and washed successively with amixture of ethanol and water (2:1), ethanol and ether and then dried toafford the title compound (45 mg) as colorless powder. The meltingpoint, infrared spectrum, NMR spectrum, elemental analysis and x-raypowder diffraction pattern of this product were identical with those ofthe compound obtained in Example 2.

Example 4

[0082](1R,5S,6S)-2-[(2S,4S)-2-[(3S)-3-(2-Guanidinoacetylamino)pyrrolidin-1-ylcarbonyl]-1-methylpyrrolidin-4-ylthio]-6-[(1R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylicAcid

[0083] (1)(1R,5S,6S)-2-[(2S,4S)-2-[(3S)-3-(2-Guanidinoacetylamino]pyrrolidin-1-ylcarbonyl]-1-methylpyrrolidin-4-ylthio]-6-[(1R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylicacid·½ ethanol (680 mg) was dissolved in water (35 ml). The mixture wasfiltered through a membrane filter. The filtrate was concentrated toapproximately 3 ml in vacuo. The resulting concentrate was allowed tostand at 0° C. overnight. The precipitated crystals were filtered andwashed with a small amount of water and then dried to afford the titlecompound as colorless crystals (294 mg).

[0084] Melting point: 235-250 (dec)

[0085] Infrared spectrum (KBr)ν max cm⁻¹: 3327, 3177, 3068, 2970, 2904,2880, 2820, 1751, 1681, 1654, 1629, 1594, 1572, 1536, 1481, 1440, 1423,1382, 1336, 1314, 1286, 1264. NMR spectrum (400 MHz, D₂O) δ ppm: 1.20(3H, dd, J=7.2, 2.2 Hz), 1.30 (3H, d, J=6.4 Hz), 1.57-1.72 (1H, m),1.93-2.13 (1H, m), 2.15-2.35 (1H, m), 2.27,2.29 (3H, s×2), 2.68-2.88(2H, m), 3.09 (1H, d, J=10.5 Hz), 3.29-3.73 (6H, m), 3.75-3.93 (2H, m),4.01 (2H, s), 4.16-4.31 (2H, m), 4.37-4.49 (1H, m).

[0086] Elemental analysis: calculated for C₂₃H₃₅N₇O₆S Cal.: C 51.38%; H6.56%; N 18.24%; S 5.96%; Found: C 51.14%; H 6.85%; N 18.26%; S 6.04%

[0087] A X-ray powder diffraction pattern of the crystalline productshown in FIG. 3 was obtained with a Cu K_(α) irradiation of λ=1.54 Å.The vertical axis of the x-ray powder diffraction pattern indicates thediffraction intensity in units of counts/second (CPS). The horizontalaxis indicates the diffraction angle as the value 2θ. The interplanarspacing d can be calculated using the equation 2d sin θ=nλ in which n is1.

[0088] (2) A similar procedure to that described above was carried outon the same scale as Example 3. To the resulting concentrate (about 1ml) was added a small amount of the colorless crystals prepared asdescribed in Example 4 (1) and the resulting mixture was allowed tostand at 0° C. overnight. The precipitated crystals were collected byfiltration and washed with a small amount of water to afford the titlecompound (20 mg).

Example 5

[0089] (1R,5S,6S)-2-[(2S,4S)-2-[(3S)-3-(2-Guanidinoacetylamino)pyrrolidin-1-ylcarbonyl]-1-methylpyrrolidin-4-ylthio]-6-[(1R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate·¼Ethanol·{fraction (3/2)} Hydrate

[0090] To a solution of 4-nitrobenzyl(1R,5S,6S)-6-[(1R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-1-methyl-2-[(3S)-3-[2-[3-(4-nitrobenzyloxycarbonyl)guanidino]-acetylamino]pyrrolidin-1-ylcarbonyl]pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate(220 g) in a mixture of tetrahydrofuran (2200 ml) and water (2200 ml)was added 7.5% palladium on carbon (220 g, which contains water (53.1%))and the resulting mixture was stirred under a hydrogen atmosphere at 30°C. for 2 hours. At the end of this time the catalyst was removed byfiltration and the filtrate was washed with ethyl acetate and wasfiltered through a membrane filter. The filtrate was concentrated toapproximately 600 ml. To the concentrate was added ethanol (1800 ml) andthe resulting mixture was stirred until crystals were precipitated, andthen this allowed to stand in an ice bath. The precipitated crystalswere filtered and washed successively with a mixture of ethanol andwater (3:1) and ethanol and then dried to afford the title compound (40g) as colorless crystals.

[0091] Melting point: 226-245° C. (dec)

[0092] Infrared spectrum (KBr)ν max cm⁻¹:3409, 3345, 3275, 3185, 2967,2884, 1761, 1674, 1644, 1586,1551, 1452, 1415, 1380, 1369, 1340, 1282,1254.

[0093] NMR spectrum (400 MHz, D₂O) δ ppm: 1.17-1.21 (4.7H, m), 1.30 (3H,d, J=6.4 Hz), 1.57-1.70 (1H, m), 1.95-2.08 (1H, m), 2.18-2.31 (1H, m),2.27, 2.29, (3H, s×2), 2.70-2.87 (2H, m), 3.08 (1H, d, J=10.8 Hz),3.31-3.72 (7H, m), 3.76-3.92 (2H, m), 4.00 (2H, S), 4.18-4.28 (2H, m),4.39-4.48 (1H, m).

[0094] Elemental analysis : calculated for C₂₃H₃₅N₇O₆S·¼C₂H₆O·{fraction(3/2)}H₂O Cal.: C 48.99%; H 6.91%; N 17.02%; S 5.56%;

[0095] Found: C 48.35%; H 6.47%; N 17.23%; S 5.67%

[0096] A X-ray powder diffraction pattern of the crystalline productshown in FIG. 4 was obtained with a Cu K_(α) irradiation of λ=1.54 Å.The vertical axis of the x-ray powder diffraction pattern indicates thediffraction intensity in units of counts/second (CPS). The horizontalaxis indicates the diffraction angle as the value 2θ. The interplanarspacing d can be calculated using the equation 2d sin θ=nλ in which n is1.

Reference Example 1

[0097] 4-Nitrobenzyl(1R,5S,6S)-2-[(2S,4S)-2-[(3S)-3-[2-[2,3-bis(4-nitrobenzyloxy-carbonyl)guanidino]-acetylamino]pyrrolidin-1-ylcarbonyl]-1-methylpyrrolidin-4-ylthio]-6-[(1R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate

[0098] Hydrazine acetate (652 mg) was added to a solution of(2S,4S)-4-acetylthio-2-[(3S)-3-[2-[2,3-bis(4-nitrobenzyloxycarbonyl)guanidino]acetylamino]pyrrolidin-1-ylcarbonyl]-1-methylpyrrolidine(4.3 g) in N,N-dimethylformamide (86 ml) and stirred at room temperaturefor 4 hours. To the resulting mixture was added 4-nitrobenzyl(1R,5R,6S)-6-[(1R)-1-hydroxyethyl]-1-methyl-2-diphenylphosphoryloxy-1-carbapen-2-em-3-carboxylate(3.53 g) and N,N-diisopropylethylamine (1.34 ml) and allowed to react at−30° C. for 3 days. At the end of this time, to the reaction mixture wasadded 1% aqueous sodium hydrogencarbonate solution and the resultingprecipitate was filtered, washed with water, and dissolved in a mixtureof tetrahydrofuran and ethyl acetate (3:7). The resulting solution waswashed successively with saturated aqueous sodium hydrogencarbonatesolution, water and saturated aqueous sodium chloride solution, driedover anhydrous magnesium sulfate and concentrated in vacuo. Theresulting residue was chromatographed on a silica gel column using 10%methanol/ethyl acetate and 20% methanol/ethyl acetate as the eluant toafford the crude desired product. The product was dissolved intetrahydrofuran and reprecipitated with a mixture of ethyl acetate andether (1:1) to give the desired compound (4.02 g) as a pale yellowpowder. Infrared spectrum (KBr)ν max cm⁻¹: 3336, 1772, 1741, 1688, 1643,1610, 1522, 1447, 1378, 1347.

[0099] NMR spectrum (270 MHz, CDCl₃) δ ppm: 1.17-1.40 (6H, m), 1.64-2.40(4H, m), 2.33 (3H, s), 2.47-2.80 (2H, m), 3.00-3.38 (3H, m), 3.46-3.83(5H, m), 3.93-4.60 (5H, m), 5.12-5.54 (6H, m), 7.21 (1H, d, J=6.5 Hz),7.46-7.70 (6H, m), 8.10-8.28 (6H, m), 8.80-9.10 (1H, br), 11.60 (1H,br).

Reference Example 2

[0100] 4-Nitrobenzyl(1R,5S,6S)-6-[(1R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-[(3S)-3-[2-[3-(4-nitrobenzyloxycarbonyl)guanidino]acetylamino]pyrrolidin-1-ylcarbonyl]-1-methylpyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate

[0101] (1) A solution of 28% sodium methylate in methanol (0.5 ml) wasadded to a solution of(2S,4S)-4-acetylthio-1-methyl-2-[(3S)-3-[2-[3-(4-nitrobenzyloxy-carbonyl)guanidino]acetylamino]pyrrolidin-1-ylcarbonyl]pyrrolidine(1.5 g) in methanol (30 ml) and stirred at room temperature for 1 hour.At the end of this time, to the resulting mixture was added INhydrochloric acid (2.73 ml) and concentrated in vacuo. The resultingresidue was chromatographed on a reverse phase column (Cosmosil 75C18PREP (trade mark) manufactured by Nacalai tesque Inc.) using a mixtureof acetonitrile and water as the eluant. The fractions containing thedesired product were combined and concentrated in vacuo. The residue waspowdered in a mixture of ethyl acetate and isopropyl ether. Powdery(2S,4S)-4-mercapto-1-methyl-2-[(3S)-3[2-[3-(4-nitrobenzyloxycarbonyl)guanidino]acetylamino]pyrrolidin-1-yl-carbonyl]pyrrolidine(806 mg) was obtained by filtration.

[0102] Infrared spectrum (KBr)ν max cm⁻¹: 3391, 3307, 3112, 3078, 2949,2877, 2786, 1732, 1639, 1548, 1522, 1448, 1380, 1347, 1291, 1211, 1154,1109.

[0103] NMR spectrum (400 MHz, CDCl₃) δ ppm: 1.64-2.15 (3H, m), 2.20-2.86(5H, m), 2.93-3.93 (9H, m), 4.16-4.35 (1H, m), 5.12 (2H, s), 6.70-6.90(1H, br), 7.00-7.85 (1H, br), 7.59 (2H, d, J=8.5 Hz), 8.23 (2H, d, J=8.5Hz), 8.18-8.40 (1H, br).

[0104] (2) N,N-Diisopropylethylamine (0.17 ml) and 4-nitrobenzyl(1R,5R,6S)-6-[(1R)-1-hydroxyethyl]-1-methyl-2-diphenylphosphoryloxy-1-carbapen-2-em-3-carboxylate(585 mg) were added to a solution of the compound (500 mg) obtained inReference example 2 (1) in N,N-dimethylformamide (5 ml) in an ice bathand allowed to react at 0° C. overnight. At the end of this time, ethylacetate and tetrahydrofuran were added to the reaction mixture, theresulting mixture was washed with 10% aqueous sodium chloride solutionand concentrated in vacuo. The resulting residue was chromatographed ona reverse phase column (Cosmosil 75C 18PREP (trade mark) manufactured byNacalai tesque Inc.) using a mixture of acetonitrile and water as theeluant. The fractions containing the desired product were combined andconcentrated in vacuo. The resulting reside was powdered in isopropylether and filtered to afford the desired compound (524 mg) as a paleyellow powder.

[0105] Infrared spectrum (KBr) ν max cm⁻¹: 3384, 3113, 3080, 2970,2875,2789, 1770, 1643, 1609, 1522,1450, 1379, 1346, 1322, 1287, 1209,1181, 1136, 1109.

[0106] NMR spectrum (400 MHz, CDCl₃) δ ppm: 1.08-2.22 (6H, m), 1.75-2.26(6H, m), 2.44-2.76 (2H, m), 2.89-3.00 (1H, m), 3.03-3.15 (1H, m),3.18-3.65 (6H, m), 3.68-3.90 (3H, m), 3.93-4.06 (1H, m), 4.13-4.35 (2H,m), 5.05-5.15 (2H, m), 5.30, 5.45 (each 1H, d, J=14.1), 7.58 (2H, dd,J=8.8, 2.7 Hz), 7.74 (2H, d, J=8.7 Hz), 8.18-8.33 (4H, m).

Reference Example 3

[0107] 4-Nitrobenzyl(1R,5S,6S)-6-[(1R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-[(3S)-3-[2-[3-(4-nitrobenzyloxycarbonyl)guanidino]acetylamino]pyrrolidin-1-ylcarbonyl]-1-methylpyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate

[0108] To a solution of(2S,4S)-4-acetylthio-1-methyl-2-[(3S)-3-[2-[3-(4-nitrobenzyloxy-carbonyl)guanidino]acetylamino]pyrrolidin-1-ylcarbonyl]pyrrolidine(500 mg) in ethanol (5 ml) was added 4N hydrogen chloride/ethyl acetate(2.7 ml) and stirred at 50° C. for 3 hours. At the end of this time,ether was added to the reaction mixture. The resulting precipitate wasseparated by decantation and dried in vacuo. To a solution of theprecipitate in N,N-dimethylformamide (10 ml) were addedN,N-diisopropylethylamine (0.63 ml) and 4-nitrobenzyl(1R,5R,6S)-6-[(1R)-1-hydroxyethyl]-1-methyl-2-diphenylphosphoryloxy-1-carbapen-2-em-3-carboxylate(541 mg) in an ice bath and allowed to react overnight. At the end ofthis time, to the reaction mixture was added 1% aqueous sodiumhydrogencarbonate solution. The resulting precipitate was filtered,washed with water and dried. The crude powder was chromatographed on asilica gel column using 30% methanol/ethyl acetate and 50%methanol/ethyl acetate as the eluant to give the desired compound (446mg). The Infrared and NMR spectra of this compound were identical withthose of the compound obtained in Reference example 2 (2).

Reference Example 4

[0109] 4-Nitrobenzyl(1R,5S,6S)-6-[(1R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-[(3S)-3-[2-[3-(4-nitrobenzyloxycarbonyl)guanidino]acetylamino]pyrrolidin-1-ylcarbonyl]-1-methylpyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate

[0110] To a solution of(2S,4S)-4-acetylthio-1-methyl-2-[(3S)-3-[2-[3-(4-nitrobenzyloxy-carbonyl)guanidino]acetylamino]pyrrolidin-1-ylcarbonyl]pyrrolidine(1.00 g) in methanol (20 ml) was added sodium methylate (98.3 mg) at 0°C. and stirred for 1 hour. At the end of this time, to the reactionmixture was added 4N hydrogen chloride/ethyl acetate (0.46 ml) andconcentrated in vacuo. In an ice bath to a solution of the resultingresidue in N,N-dimethylformamide (10 ml) were added a solution ofN,N-diisopropylethylamine (0.32 ml) and 4-nitrobenzyl(1R,5R,6S)-6-[(1R)-1-hydroxyethyl]-1-methyl-2-diphenylphosphoryloxy-1-carbapen-2-em-3-carboxylate(1.08 g) in N,N-dimethylformamide and allowed to stand at 0° C.overnight. At the end of this time, to the reaction mixture was added 1%aqueous sodium hydrogencarbonate solution. The resulting precipitate wasfiltered, washed with water and dried. The crude powder waschromatograhed on a silica gel column using methanol/ethyl acetate=⅓ andmethanol/ethyl acetate=½ as the eluant to give the desired compound (975mg). The Infrared and NMR spectra of this compound were identical withthose of the compound obtained in Reference example 2-(2).

Test Example 1

[0111] Stability test

[0112] The crystalline compounds obtained in Example 1, 2, 4 and 5 werekept for about 2 months in a desiccator at 40° C. and 75% relativehumidity and in a desiccator at 60° C. in which silica gel was placed,respectively. The non-crystalline powdered compound (lyophilizedproduct) obtained according to the procedure described in U.S. Pat. No.6,090,802 Example 72 and in Japanese Patent Application Publication No.Hei-11-071277, which is a reference sample, was kept under the sameconditions described above. The remaining amount of the crystallinecompounds and the non-crystalline powdered compound were determinedafter 7, 20, 21, 28, and 56 days by high pressure liquid chromatographyon a L-Column ODS (4.6 mm Φ×150 mm, (trade mark) manufactured byKagakuhinn kennsa kyoukai) eluting with 20 mM KH₂PO₄ (pH 7.0):CH₃CN=96:4 at 1.0 ml/minute at 60° C. using an ultraviolet wave lengthof 300 nm. The remaining percentages of these compounds were calculatedfrom the remaining amount of them and show in Tables 1-4. TABLE 1Stability in desiccator at 40° C. and 75% relative humidity Remainingpercentage of compound Compound After 20 days After 56 days Crystalsobtained in Example 1 98.5 92.9 Crystals obtained in Example 2 95.8 87.3Crystals obtained in Example 4 99.4 94.3 Non-crystalline powder 30.2 0.3 (lyophilized product)

[0113] TABLE 2 Stability in desiccator at 60° C. in which silica gel wasplaced Remaining percentage of compound Compound After 21 days After 56days Crystals obtained in Example 1 98.5 96.4 Crystals obtained inExample 2 100.1 96.8 Crystals obtained in Example 4 100.4 100.9Non-crystalline powder 74.1 56.5 (lyophilized product)

[0114] TABLE 3 Stability in desiccator at 40° C. and 75% relativehumidity Remaining percentage of compound Compound After 7 days After 28days After 56 days Crystals obtained in 98.9 94.6 83.2 Example 5

[0115] TABLE 4 Stability in desiccator at 60° C. in which silica gel wasplaced Remaining percentage of compound Compound After 7 days After 28days After 56 days Crystals obtained in 98.3 97.0 97.6 Example 5

[0116] From the Tables 1 and 3, it is clear that the non-crystallinepowder of compound (I) (lyophilized product) was very unstable at 40° C.and 75% relative humidity, that is, after 20 days the percentage ofremaining amount of the compound is 30.2% and after 56 days it is only0.3%, and that the percentages of remaining amount of the crystallinecompounds of this invention are more than 83, respectively under thesame conditions.

[0117] From the Tables 2 and 4, it is clear that the non-crystallinepowder of compound (I) (lyophilized product) was also unstable at 60° C.under dry conditions, that is, after 21 days the percentage of remainingamount of the compound is 74.1% and after 56 days that is 56.5%, andthat the percentages of remaining amount of the crystalline compounds ofthis invention are more than 96, respectively under the same conditions.

[0118] These results show that the crystalline compounds of thisinvention are extremely stable compared to the non-crystalline powder(lyophilized product) belonging to the previous invention.

Test Example 2

[0119] Antibiotic Activity Test

[0120] The MIC (μg/ml), the lowest concentration of antibiotic whichinhibits growth of the test bacterial strain, was determined by theagar-plate dilution method. The crystalline compounds obtained inExample 1, 2, 4 and 5 of this invention were evaluated against variousbacterial strains by determining the MIC of each compound with respectto each strain. Table 5 illustrates the result of such experiments.TABLE 5 Antibiotic activity (MIC, μg/ml) Crystalline compounds obtainedin Bacterial strain Example 1 2 4 5 Staphylococcus aureus 209P ≦0.01≦0.01 ≦0.01 ≦0.01 S. aureus 535 (MRSA) 1.5 1.5 1.5 1.5 Escherichia coliNIHJ ≦0.01 ≦0.01 ≦0.01 ≦0.01 E. coli 609 0.02 ≦0.01 0.02 ≦0.01Klebisella pneumoniae 806 ≦0.01 ≦0.01 ≦0.01 ≦0.01 K. pneumoniae 846≦0.01 ≦0.01 ≦0.01 ≦0.01 Enterobacter cloacae 963 0.05 0.05 0.05 0.05Serratia marcescens 1184 ≦0.01 ≦0.01 ≦0.01 ≦0.01 Pseudomonas aeruginosa1001 0.2 0.2 0.2 0.39

Formulation Example 1

[0121] Injections

[0122] The crystalline compound obtained in Example 1 (250 mg) is usedto fill a vial and shielded with a stopper under sterile conditions.Pharmaceutical additives known to those skilled in the art such as alocal anaesthetic agent, for example, lidocaine hydrochloride can beadded to the vial, if necessary. The sterile solid compositions can bedissolved in an injectable medium such as water for injectionimmediately before use.

BRIEF DESCRIPTION OF THE DRAWINGS

[0123]FIG. 1 shows the powder diffraction pattern of crystalline(1R,5S,6S)-2-[(2S,4S)-2-[(3S)-3-(2-guanidinoacetylamino)pyrrolidin-1-ylcarbonyl]-1-methylpyrrolidin-4-ylthio]-6-[(1R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic Acid·½Carbonate·½ Ethanol (I-1).

[0124] The diffraction pattern was obtained with a Cu K_(α) irradiationof λ=1.54 Å to the crystals. The vertical axis of the x-ray powderdiffraction pattern indicates the diffraction intensity in units ofcounts/second (CPS). The horizontal axis indicates diffraction angle asthe value 2θ.

[0125]FIG. 2 shows the x-ray powder diffraction pattern of crystalline(1R,5S,6S)-2-[(2S,4S)-2-[(3S)-3-(2-guanidinoacetylamino)pyrrolidin-1-ylcarbonyl]-1-methyl-pyrrolidin-4-ylthiol-6-[(1R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylicacid·½ ethanol (I-2).

[0126] The diffraction pattern was obtained with a Cu K_(α) irradiationof λ=1.54 Å to the crystals. The vertical axis of the x-ray powderdiffraction pattern indicates the diffraction intensity in units ofcounts/second (CPS). The horizontal axis indicates diffraction angle asthe value 2θ.

[0127]FIG. 3 shows the x-ray powder diffraction pattern of(1R,5S,6S)-2-[(2S,4S)-2-[(3S)-3-(2-guanidinoacetylamino)pyrrolidin-1-ylcarbonyl]-1-methylpyrrolidin-4-ylthio]-6-[(1R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid (I).

[0128] The diffraction pattern was obtained with a Cu K_(α) irradiationof λ=1.54 Å to the compound (I). The vertical axis of the x-ray powderdiffraction pattern indicates the diffraction intensity in units ofcounts/second (CPS). The horizontal axis indicates diffraction angle asthe value 2θ.

[0129]FIG. 4 shows the x-ray powder diffraction pattern of crystalline(1R,5S,6S)-2-[(2S,4S)-2-[(3S)-3-(2-guanidinoacetylamino)pyrrolidin-1-ylcarbonyl]-1-methyl-pyrrolidin-4-ylthio]-6-[(lR)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid·¼ethanol·{fraction (3/2)} hydrate (I-3).

[0130] The diffraction pattern was obtained with a Cu K_(α) irradiationof λ=1.54 Å to the crystals. The vertical axis of the x-ray powderdiffraction pattern indicates the diffraction intensity in units ofcounts/second (CPS). The horizontal axis indicates diffraction angle asthe value 2θ.

What is claimed is:
 1. A 1-methylcarbapenem derivative of formula (I) ora pharmaceutically acceptable salt thereof in crystalline form:


2. A 1-methylcarbapenem derivative of formula (I-1) in crystalline form:


3. A 1-methylcarbapenem derivative of formula (I-2) in crystalline form:


4. A 1-methylcarbapenem derivative of formula (I) in crystalline form:


5. A 1-methylcarbapenem derivative of formula (I-3) in crystalline form:


6. A pharmaceutical composition for the prevention or treatment ofbacterial infections comprising a therapeutically effective amount of a1-methylcarbapenem derivative of formula (I) or a pharmaceuticallyacceptable salt thereof in crystalline form as claimed in claim 1 as anactive ingredient together with a diluent or carrier therefor.
 7. Apharmaceutical composition for the prevention or treatment of bacterialinfections comprising a therapeutically effective amount of a1-methylcarbapenem derivative of formula (I-1) in crystalline form asclaimed in claim 2 as an active ingredient together with a diluent orcarrier therefor.
 8. A pharmaceutical composition for the prevention ortreatment of bacterial infections comprising a therapeutically effectiveamount of a 1-methylcarbapenem derivative of formula (I-2) incrystalline form as claimed in claim 3 as an active ingredient togetherwith a diluent or carrier therefor.
 9. A pharmaceutical composition forthe prevention or treatment of bacterial infections comprising atherapeutically effective amount of a 1-methylcarbapenem derivative offormula (I) in crystalline form as claimed in claim 4 as an activeingredient together with a diluent or carrier therefor.
 10. Apharmaceutical composition for the prevention or treatment of bacterialinfections comprising a therapeutically effective amount of a1-methylcarbapenem derivative of formula (I-3) in crystalline form asclaimed in claim 5 as an active ingredient together with a diluent orcarrier therefor.
 11. A method for preventing or treating bacterialinfections which comprises administering to a warm-blooded animal inneed of such prevention or treatment a pharmacologically effectiveamount of a 1-methylcarbapenem derivative of formula (I) or apharmaceutically acceptable salt thereof in crystalline form as claimedin claim
 1. 12. A method for preventing or treating bacterial infectionswhich comprises administering to a warm-blooded animal in need of suchprevention or treatment a pharmacologically effective amount of a1-methylcarbapenem derivative of formula (I-1) in crystalline form asclaimed in claim
 2. 13. A method for preventing or treating bacterialinfections which comprises administering to a warm-blooded animal inneed of such prevention or treatment a pharmacologically effectiveamount of a 1-methylcarbapenem derivative of formula (I-2) incrystalline form as claimed in claim
 3. 14. A method for preventing ortreating bacterial infections which comprises administering to awarm-blooded animal in need of such prevention or treatment apharmacologically effective amount of a 1-methylcarbapenem derivative offormula (I) in crystalline form as claimed in claim
 4. 15. A method forpreventing or treating bacterial infections which comprisesadministering to a warm-blooded animal in need of such prevention ortreatment a pharmacologically effective amount of a 1-methylcarbapenemderivative of formula (I-3) in crystalline form as claimed in claim 5.